Susceptibility gene region closely related to psoriasis and method for detecting psoriasis susceptible population
A susceptibility gene, susceptible population technology, applied in the fields of molecular genetics and dermatology, can solve problems such as lack of identification methods
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Embodiment 1
[0030] The following uses the MassARRAY system (sequenom, Inc) to detect the LCE gene SNP difference in this Block region and the haplotype difference derived therefrom to determine the susceptible population of psoriasis.
[0031] 1. Extraction of genomic DNA
[0032]The subjects all draw 5ml of anticoagulant blood from the peripheral vein, and use the DNA extraction kit to extract genomic DNA. The steps are as follows: ① Pipette 1.0ml cell lysate (FG1) into a labeled 2.0ml centrifuge tube, take 400ul blood sample into the corresponding centrifuge tube, invert and mix 20-30 times until no blood clots appear, centrifuge at 16,000g 4 minutes. ②Carefully pour the supernatant, drain on absorbent paper for 30 seconds, add 200ul of denaturation buffer / proteinase K (FG2 / PK) solution, shake and mix each part immediately until no cell clumps appear, wait until all of them are added. Check carefully for incompletely lysed samples. If so, it should be shaken thoroughly. ③After a short centr...
Embodiment 2
[0073] Next, the differences in the SNP sites of the LCE gene in this Block region are detected by gene sequencing to determine the susceptible population of psoriasis.
[0074] 1. Extraction of genomic DNA
[0075] After extracting anticoagulant blood from the peripheral vein of the subject, genomic DNA is extracted. The steps are shown in the first part of the first implementation method.
[0076] 2. PCR amplification of target fragments containing SNP sites
[0077] Design the upstream and downstream primers that can amplify the SNP site of the LCE gene in this Block region, and configure it as follows
[0078]
Concentration
Volume
(μL)
HPLC grade water
N / A
1.750
10X PCR buffer containing 15mM MgCl2
1.25x(1.875mM MgCl2)
0.625
25mM MgCl2
1.625mM
0.325
25mM dNTP mix
500uM
0.100
0.5uM primer
0.1uM
1.000
5U / μl HotStar DNA polymerase
1Unit
0.200
...
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